Abstract: OBJECTIVE To explore the impact of environmental cleaning and disinfection on prevalence of nosocomial infection in intensive care unit (ICU) so as to provide guidance for prevention of the nosocomial infection in ICU. METHODS The patients who were hospitalized in the ICUs of Huangshi Central Hospital of Edong Healthcare Group from Jan 1, 2016 to Oct 31, 2016 and from Jan 1, 2017 to Oct 31,2017 were enrolled in the study and divided into the original disinfection mode group (wiping with disinfectant containing chlorine) and the new disinfection mode group (wiping with disposable sterilizing wet wipe). The qualified rate of hygienic sampling and incidence of infections in the ICUs were observed and compared between the two groups. RESULTS The positive rate of hygienic sampling was 83.33% in the original disinfection mode group, 98.61% in the new disinfection mode group (P＜0.05); the incidence of infections in the ICU was 10.19% in the original disinfection mode group, 3.38% in the new disinfection mode group (P＜0.05). The incidence rate of ventilator-associated pneumonia (VAP) was 8.39‰ in the original disinfection mode group, 3.80‰ in the new disinfection mode group (P＜0.05); the incidence rate of central line associated-bloodstream infection (CLABSI) was 0.86‰ in the original disinfection mode group, 0‰ in the new disinfection mode group; the incidence rate of catheter-associated urinary tract infection (CAUTI) was 1.20‰ in the original disinfection mode group, 1.24‰ in the new disinfection mode group; the proportion of multidrug-resistant strains causing nosocomial infection was 19.48% in the original disinfection mode group, 7.95% in the new disinfection mode group (P＜0.05). CONCLUSION The improved cleaning and disinfection mode may raise the quality of cleaning and disinfection in the ICU, prevent and reduce the incidence of nosocomial infection and VAP as well as the proportion of multidrug-resistant strains causing the nosocomial infection.